Project Title: Bio-nanotechnology approach for treatment of lung cancer Project Summary Lung cancer is the leading cause of cancer-related deaths worldwide. Non-Small Cell Lung Carcinoma (NSCLC) is the most common type of lung cancer, accounting for more than 80% of all lung cancer cases. Chemotherapy is the primary pre-operative and post-operative treatment of NSCLC. However, the efficiency of chemotherapy remains relatively low in most patients and is limited by insufficient specificity, low drug accumulation, and retention in the lungs with severe adverse side effects of treatment. Recently, small molecule Tyrosine Kinase (TK) inhibitors which act on the Epidermal Growth Factor Receptors (EGFRs) were introduced for treatment of NSCLC. However, even the latest generation of EGFR inhibitors cause severe systemic toxicities and are ineffective in preventing non-canonical EGFR signaling. As a result, only approximately 10% of patients with NSCLC benefit from this therapy. In order to overcome these limitations, a novel multi-tier biotechnology treatment approach is proposed that includes: (1) suppression of all four types of EGFR-TKs by a pool of small interfering RNAs (siRNAs); (2) induction of cell death by an anticancer drug, (3) enhancing the efficiency of the treatment by the local inhalatory delivery of therapeutic agents to the lungs (passive targeting), (4) active receptor-mediated targeting of the therapy specifically to cancer cells and (5) increasing the stability, solubility, and cellular penetration of siRNA and drug by using Nanostructured Lipid Carriers (NLC). We hypothesize that the application of this nanotechnology-based tumor-targeted, multifunctional approach will substantially enhance the efficiency of the treatment of NSCLC and reduce adverse side effects of chemotherapy. The main objective of the current research is to test the stated hypothesis and develop a novel nanoscale-based technology to carry out proof-of-concept of the proposed strategy. The specific aims of the proposed research are: (1) to engineer, synthesize, and characterize a multifunctional, multicomponent Delivery System (DS) containing NLC, a pool of siRNAs targeted to EGFR-TKs, paclitaxel (TAX) as an anticancer drug, and a Luteinizing Hormone-Releasing Hormone (LHRH) peptide as a targeting moiety specific to receptors overexpressed in lung cancer cells; (2) to examine efficiency of active (LHRH receptor-mediated) and passive (local inhalation delivery) dual tumor targeting of NLC-based DS; (3) to characterize the efficiency of combinatorial gene and chemotherapy for silencing of EGFR-TK signaling pathways and cell death induction; (4) to evaluate in vivo antitumor activity and adverse side effects of targeted and non-targeted NLC-based DS in clinically relevant orthotopic mouse models of primary human NSCLC xenografts. It is expected that the proposed approach and the use of the developed multifunctional NLC-based DS will substantially enhance the efficiency of therapy of NSCLC and limit adverse side effects of the treatment. Experimental data obtained will provide the proof-of-concept of the proposed approach, and can potentially make a considerable impact on the field of drug delivery and improve the efficiency of therapy for lung and other types of cancer.